FISH on sperm: spot-counting to stop counting? Not yet.
OBJECTIVE
To evaluate the reliability and applicability of the spot-counting system (Cytovision Spot AX workstation) which offers an alternative to the tedious manual analysis of sperm fluorescence in situ hybridization (FISH).


METHODS
Manual and automatic analyses were performed and compared.


METHODS
Universitat Autònoma de Barcelona.


METHODS
Twenty-four men who asked for information on infertility showing different seminal parameters.


METHODS
A semen sample for each patient was collected and prepared for FISH.


METHODS
A dual-color FISH using specific probes for chromosomes 13 and 21 and a triple-color FISH with centromeric probes for chromosomes 18, X, and Y were used (Vysis). Standard FISH analysis was carried out. Automatic analysis was subsequently performed using a Spot AX system.


RESULTS
Overall, we performed 120 comparisons. In 116 out of 120 (96.67%), the percentage of anomalies reported using manual counting fell within the incidence detected using the automatic system. In the remaining comparisons, statistical differences were detected (4 out of 120; 3.33%). Time consumed by the automatic analysis was always higher than the manual one, being influenced by the characteristics of the preparations.


CONCLUSIONS
The spot-counting system has potential, but before the service is ready to be offered, we still need to overcome some limitations associated with it.
Molina
Òscar
Ò
Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain.
Sarrate
Zaida
Z
Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain.
Vidal
Francesca
F
Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain.
Blanco
Joan
J
Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain. Electronic address: joan.blanco@uab.cat.
D002882:Chromosomes, Human, Pair 13; D002887:Chromosomes, Human, Pair 18; D002891:Chromosomes, Human, Pair 21; D004171:Diploidy; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D008297:Male; D015203:Reproducibility of Results; D012588:Scintillation Counting / Q000295:instrumentation / Q000379:methods*; D055101:Semen Analysis / Q000379:methods*; D012730:Sex Chromosomes; D013094:Spermatozoa / Q000648:ultrastructure; D024182:Uniparental Disomy / Q000175:diagnosis